Practical Synthesis of Isomerically Pure 5-and 6-Carboxytetramethylrhodamines, Useful Dyes for DNA Probes
Practical Synthesis of Isomerically Pure 5-and 6-Carboxytetramethylrhodamines, Useful Dyes for DNA Probes
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DOI：
10.1021/bc900037b
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发表时间：
2009-08-01
影响因子：
4.7
通讯作者：
Shmanai, Vadim V.
中科院分区：
文献类型：
--
作者：
Kvach, Maksim V.;Stepanova, Irina A.;Shmanai, Vadim V.
报道了5-和6-羧基四甲基罗丹明（TAMRAs）的简易合成方法。3-二甲氨基苯酚与1,2,4-苯三羧酸甲酰化得到4-二甲氨基-2 -羟基-2',4'(5')-二羧基二苯甲酮的混合物，在甲醇和乙酸中重结晶后很容易分离成单独的化合物。单个二苯酮与3-二甲氨基苯酚反应得到5-或6-羧基四甲基罗丹明。将染料转化为适合于合成寡核苷酸的羟脯氨酸基磷酰胺试剂。寡核苷酸上的5-和6-TAMRA异构体表现出相似的吸收和发射光谱。染料的荧光量子产率与寡核苷酸序列邻区是否存在dG核苷有关。几种能量转移引物在其5'端含有（6-FAM）dT(n)（6-TAMRA）染料体系（n = 0,2）。制备了4、6、8、10、12、14)，并对其光谱性质进行了研究。
Simple and scalable synthesis of 5- and 6-carboxytetramethylrhodamines (TAMRAs) is reported. Acylation of 3-dimethylaminophenol with 1,2,4-benzenetricarboxylic anhydride afforded a mixture of 4-dimethylamino-2hydroxy-2',4'(5')-dicarboxybenzophenones, which can be easily separated into individual compounds upon recrystallization from methanol and acetic acid. Individual benzophenones were reacted with 3-dimethylaminophenol to give 5- or 6-carboxytetramethylrhodamines. The dyes were converted into hydroxyprolinol-based phosphoramidite reagents suitable for oligonucleotide synthesis. 5- and 6-TAMRA isomers on oligonucleotides showed similar absorption and emission spectra. Fluorescence quantum yield of the dyes correlates with the presence of dG nucleosides in the adjacent region of oligonucleotide sequence. Several energy transfer primers containing on their 5'-termini (6-FAM)dT(n)(6-TAMRA) dye system (n = 0, 2. 4, 6, 8, 10, 12, 14) were prepared, and their spectral properties were studied.